A green leaf volatile, (Z)-3-hexenyl-acetate, mediates differential oviposition by Spodoptera frugiperda on maize and rice.

BACKGROUND: Insects rely on chemosensory perception, mainly olfaction, for the location of mates, food sources, and oviposition sites. Plant-released volatile compounds guide herbivorous insects to search for and locate their host plants, further helping them to identify suitable positions for oviposition. The fall armyworm Spodoptera frugiperda (S. frugiperda) was found to invade China in 2019 and has since seriously threatened multiple crops, particularly maize and rice. However, the chemical and molecular mechanisms underlying oviposition preference in this pest are not fully understood. Here, the oviposition preference of S. frugiperda on maize and rice plants was investigated. RESULTS: GC-EAD and GC-MS/MS techniques were used to identify the antennally active volatiles from maize and rice plants. The attraction and oviposition stimulation of identified components to female adults were tested in both laboratory and field settings. The odorant receptors (ORs) on female antennae were expressed in Xenopus oocytes, and their functions evaluated by RNAi. Ten and eleven compounds of maize and rice plants, respectively, were identified to possess electrophysiological activity from headspace volatiles. Among these compounds, (Z)-3-hexenyl-acetate specifically presented in maize volatiles was found to play a critical role in attracting females and stimulating oviposition compared to rice volatiles. Among the cloned ORs on the antennae of both sexes, SfruOR23 with highly female-biased expression mediated the responses of females to (Z)-3-hexenyl-acetate. Knockdown of SfruOR23 using RNAi markedly reduced the electrophysiological response of female antennae and oviposition preference to the compound. CONCLUSIONS: (Z)-3-Hexenyl-acetate is a key volatile mediating the host and oviposition preference of S. frugiperda on maize. The olfactory receptor of (Z)-3-hexenyl-acetate was identified to be SfruOR23, which is mainly expressed in the antennae of S. frugiperda.

Structure and distribution of antennal sensilla in Pseudosymmachia flavescens (Brenske) (Coleoptera: Scarabaeidae: Melolonthinae).

Morphology, microstructure, and distribution of antennal sensilla were compared between female and male Pseudosymmachia flavescens (Brenske) (Coleoptera: Scarabaeidae: Melolonthinae). Lamellate antennae of P. flavescens were shown to have typical scape, pedicel, and flagellum segments. The flagellum consists of a four-segmented funicle and a three-segmented club. The lengths of their pedicel and funicle were found to be similar in females and males. Distinct sexual differences were observed in the length of lamellar segments. Nine types/subtypes of sensilla were identified on the antennae of both sexes, including Böhm sensilla, sensilla trichodea, sensilla basiconica (SB), and two subtypes for sensilla chaetica, sensilla coeloconica, and sensilla placodea each. Olfactory sensilla (e.g., SB and placodea) are mainly located on three lamellar segments of the antennal club. Variation was also seen in abundance of various types of antennal sensilla, with males possessing significantly more sensilla than females. Sensilla placodea were the most abundant, and their number in males was twice of that in females, showing a clear sexual dimorphism. The difference in the distribution of sensilla placodea might reflect their roles in sexual chemical communication.

Electrophysiological and Alarm Responses of Solenopsis invicta Buren (Hymenoptera: Formicidae) to 2-Ethyl-3,5-dimethylpyrazine (Short Title: EAG and Behavioral Responses of Fire Ants to Pyrazine).

2-Ethyl-3,5-dimethylpyrazine is an isomer of 2-ethyl-3,6-dimethylpyrazine, the alarm pheromone component of the red imported fire ant, Solenopsis invicta Buren. The pyrazine was synthesized and its alarm activity was investigated under laboratory conditions. It elicited significant electroantennogram (EAG) activities, and released characteristic alarm behaviors in fire ant workers. The EAG and alarm responses were both dose-dependent. Two doses of the pyrazine, 1 and 100 ng, were further subjected to bait discovery bioassays. Fire ant workers excited by the pyrazine were attracted to food baits, and their numbers increased over time. Ants displayed very similar response patterns to both low and high doses of the pyrazine. The pyrazine impregnated onto filter paper disc attracted significantly more fire ant workers than the hexane control for all observation time intervals at the low dose, and in the first 15 min period at the high dose. The pyrazine loaded onto food bait directly tended to attract more fire ant workers than the hexane control. These results support the potential use of 2-ethyl-3,5-dimethylpyrazine to enhance bait attractiveness for the control of S. invicta in invaded regions.

[Effect of electroacupuncture on muscular atrophy and Slit/Robo signaling in sciatic nerve and lumbar spinal cord tissues in rats with sciatic nerve injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Huantiao"(GB30) and" Zusanli"(ST36)on muscular atrophy and expression of Slit-Robo GTPase-activating protein(srGAP)1, 2 and 3 in the injured sciatic nerve and lumbar spinal cord tissues in sciatic nerve injury (SNI) rats, so as to reveal its mechanisms underlying improvement of peripheral nerve injury (PNI)．. METHODS: A total of 120 healthy male SD rats were randomly divided into control, sham-operation, model and EA groups (n＝30 rats in each) which were further divided into 7, 15 and 23 d subgroups (n＝10 rats in each subgroup). The SNI model was established by transecting the right sciatic nerve beneath the piriformis and immediately subsequent end-to-end suture. Rats of the sham operation group received an incision of the corresponding skin and suture. EA (5 Hz/20 Hz, 2－3 mA) was applied to the right GB30 and ST36 for 15 min, once daily, 6 days a week separately for 1，2 and 3 weeks. Rats in the sham-operation and model groups were grasped in the similar procedure as the EA group. The wet weight of gastrocnemius muscles (WWG) on both sides was measured to calculate the recovery rate (weight of the right WWG/weight of the left WWG×100%), and the expression levels of srGAP1, srGAP2 and srGAP3 proteins in the sciatic nerve and the spinal cord (L4-L6) tissues were detected by Western blot. RESULTS: After modeling and compared with the control and sham-operation groups, the recovery rate of WWG was significantly reduced, and the expression levels of srGAP1, srGAP2 and srGAP3 proteins of the sciatic nerve and lumbar spinal cord on day 7, 15 and 23 were considerably increased in the model group (P<0.01). Following the EA treatment, the reco-very rate of WWG was obviously increased and the expression levels of srGAP1, srGAP2 and srGAP3 proteins of both sciatic nerve and spinal cord on day 7, 15 and 23 were further significantly up-regulated in the EA group relevant to the model group (P<0.05，P<0.01). In addition, the expression levels of the 3 proteins in both sciatic nerve and lumbar spinal cord peaked on day 15 and attenuated on day 23. CONCLUSION: EA of GB30 and ST36 may relieve gastrocnemius atrophy in SNI rats, which is related to its function in up-regulating the Slit/Robo signaling in the sciatic nerve and lumbar spinal cord to promote the axonal targeting regeneration and repair of axonal plasma nutrition transportation.

A Practical Technique for Electrophysiologically Recording from Lamellated Antenna of Scarab Beetle.

Host-plant volatiles play a key role in finding mate and suitable host plants of phytophagous scarab beetles. Hence it is immensely important to collect and identify these volatiles. The gas chromatography coupled with electroantennographic detection (GC-EAD) technique has been used as a rapid and convenient tool for the identification of physiologically active components from plants. Here, we describe a practical method for electrophysiologically recording from lamellated antenna of scarab beetles. This method enables direct electroantennogram (EAG) recordings from antennal club without damage to the antenna in a similar manner to the conventional cut-tip EAG recording technique for clavate antenna. The headspace volatiles from walnut (Juglans regia L.) trees were collected with a Poropak-Q trap at dusk and then analyzed with GC-EAD. Those volatile compounds that elicited electrophysiological responses on the antennae of a scarab beetle, Metabolus flavescens Brenske (Coleoptera: Scarabeidae: Melolonthinae) were determined by means of gas chromatography-mass spectrometry (GC-MS). The lamella directly connected to the recording electrode was held apart from the other two lamellae on the antenna with a minuten pin and a disposable syringe needle. In order to improve electrical contact, a surfactant, Tween® 80, was used to lower the surface tension of Beadle-Ephrussi Ringer solution. This study demonstrated that addition of 0.05% Tween® 80 to the Beadle-Ephrussi Ringer solution suppressed baseline noise and assured significantly greater EAG response in general. Due to its simplicity and efficiency, this method may also be useful for studying the electrophysiology of other insect species having club-like antennae.

HPLC Separation of 2-Ethyl-5(6)-methylpyrazine and Its Electroantennogram and Alarm Activities on Fire Ants (Solenopsis invicta Buren).

2-Ethyl-3,6-dimethylpyrazine (EDMP) was an alarm pheromone component isolated from the mandibular gland of the red imported fire ant, Solenopsis invicta Buren. Several pyrazine analogues have been previously found to elicit significant alarm responses in S. invicta workers. This study aimed to separate the commercially available 2-ethyl-5(6)-methylpyrazine (EMP), i.e., a mixture of 2-ethyl-6-methylpyrazine (2E6MP) and 2-ethyl-5-methylpyrazine (2E5MP), and to examine both electroantennogram (EAG) and behavioral responses of S. invicta workers to EMP and the purified isomers. HPLC separations were achieved using a polysaccharide chiral stationary phase (Chiralpak AD-H) column with both mobile phases: Cyclohexane/isopropanol, and hexane/isopropanol. A ratio of 99:1 was selected for the separation of EMP at semipreparative level. The structures of the isomers obtained through the cyclohexane/isopropanol mobile phase were confirmed by detailed analyses of 2D-HSQC- and -HMBC-NMR data. The two isomers showed differential methine C⁻H correlations evidenced by 2D-HMBC-NMR spectra. The two concentrated fractions obtained through hexane/isopropanol mobile phase were subjected to EAG test and behavioral bioassay on S. invicta workers. The two HPLC−purified isomers, 2E6MP and 2E5MP, and their mixture (1:1) at same dose elicited similar EAG and alarm responses, indicating that these two isomers are equally active. The 2D-NMR−spectroscopic characterization, and electrophysiological and alarm activities of 2E6MP and 2E5MP were reported here for the first time.

[Histological observations of Medaka(Oryzias latipes)gonad sexual differentiation and development].

Over the past few decades, Medaka (Oryzias latipes) has become a model animal in teleost species research due both to its short reproductive cycle and efficient proliferate capacity. Unfortunately, however, systematic data of its sexual differentiation and development have yet to be obtained. In the present study, we observed Medaka gonad development from the earliest recognizable stages through differentiation to maturation under a light microscope, after paraffin sectioning and hematoxylin-eosin staining. The results showed that among juveniles aged 5 to 10 days, the gonad located on the right side of the dorsal abdominal cavity and the germ cells was significantly larger than the surrounding somatic cells. Ten days after hatching, sexual differences between females and males became obvious, and germline cysts derived from oogonium division were present in the gonad. At 50 days after hatching, ovarian cavities and mature sperm were observed. Moreover, during the study we also observed the coexistence of two sexual characteristics during a male's development. Together, these observations fill current gaps in developmental and genetic biology that will allow a more efficient use of Medaka as an experimental model.